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Topic: Western blot - pProtein v.s. Protein  (Read 2057 times)

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Offline Nescafe

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Western blot - pProtein v.s. Protein
« on: November 28, 2012, 10:24:28 PM »
Hi,

Lets say we are doing western and we are looking at the phosphorylation levels of a particular protein of XkDa. On the SDS-Gel, would not the Phosphorylated protein (pX) and the protein that is not phosphorylated run to the same point on the gel? SO they will be overlapping? Or is phosphorus enough change in charge to mass ratio to separate them on the gel?

Thanks in advance!

Nescafe.

Offline Yggdrasil

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Re: Western blot - pProtein v.s. Protein
« Reply #1 on: November 28, 2012, 10:51:08 PM »
In some cases, phosphorylation does alter the mobility of a protein on an SDS-PAGE gel, but this is not always the case and it depends on the specific protein.  In general, however, you can use phospho-specific antibodies to distinguish between the phosphorylated and unphosphorylated versions of your protein.

Offline Babcock_Hall

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Re: Western blot - pProtein v.s. Protein
« Reply #2 on: November 29, 2012, 11:19:15 AM »
I seem to recall seeing a native gel of a protein called Spo0F that separated phosphorylated versus dephosphorylated forms.

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